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. 2026 Mar 28;12:121. doi: 10.1038/s41531-026-01319-6

Fig. 2. Augmenting O-GlcNAcylation with GlcN attenuates LPS-induced neuroinflammation and dopaminergic neurodegeneration in a murine PD model.

Fig. 2

The mice received bilateral injections of LPS (5 μg) into the SN of the brain. GlcN was administered intraperitoneally at a dose of 200 mg/kg, three times per week for 4 weeks. Motor coordination and bradykinesia were evaluated using the rotarod test (A) and pole test (B) at 4 weeks post-injection (n = 6 per group). TH immunohistochemistry was performed to assess dopaminergic neurons in the SN (C) and ST (D). Scale bars = 100 μm (n = 3 per group). (E) Western blot analysis of TH expression in SN lysates. Quantification was normalized to β-actin (n = 3 per group). Representative immunofluorescence images of SN tissue showing GFAP (red), TH (green), DAPI (blue) (F) and Iba1 (red), TH (green), DAPI (blue) (G). Scale bars = 50 μm (n = 3 per group). H Western blot analysis of inflammatory markers iNOS and COX2 in SN lysates. Quantification was normalized to β-actin (n = 3 per group). I Western blot analysis of NF-κB pathway components, including phospho-p65 (p-p65), phospho-IκB (p-IκB), total p65, and total IκB in SN lysates. Quantification of p-p65 was normalized to total p65, and p-IκB was normalized to total IκB (n = 3 per group). J Western blot analysis of inflammasome-related proteins, including NLRP3, ASC, cleaved caspase-1, total caspase-1, and β-actin in SN lysates. Quantification of NLRP3 and ASC was normalized to β-actin; cleaved caspase-1 was normalized to total caspase-1 (n = 3 per group). K Representative immunofluorescence images of ASC (green), Iba1 (red), and DAPI (blue) in SN tissue. Scale bar = 10 μm (n = 3 per group). L ELISA analysis of pro-inflammatory cytokines (TNF-α, IFN-γ, IL-1β) and anti-inflammatory cytokines (IL-4, IL-10, TGF-β) in SN tissue (n = 3 per group). Data are presented as mean SEM; *p < 0.05, **p < 0.01, ***p < 0.001 versus control, #p < 0.05, ##p < 0.01, ###p < 0.001 versus LPS. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc multiple comparison test.